Tethered multifluorophore motion reveals equilibrium transition kinetics of single DNA double helices.

ABSTRACT: We describe a tethered multifluorophore motion assay based on DNA origami for revealing bimolecular reaction kinetics on the single-molecule level. Molecular binding partners may be placed at user-defined positions and in user-defined stoichiometry; and binding states are read out by tracking the motion of quickly diffusing fluorescent reporter units. Multiple dyes per reporter unit enable singe-particle observation for more than 1 hour. We applied the system to study in equilibrium reversible hybridization and dissociation of complementary DNA single strands as a function of tether length, cation concentration, and sequence. We observed up to hundreds of hybridization and dissociation events per single reactant pair and could produce cumulative statistics with tens of thousands of binding

SUBMITTER: Schickinger M 

PROVIDER: S-EPMC6094131 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature